I am trying to remove the electrostatically bound proteins using DPBS at various concentrations viz. 2X, 4X, 6X and 8x. The SDS-PAGE shows similar profile for 2X & 4X DPBS. I am unable to understand that how at nearly 600 mosm the sperm didn't burst(4xDPBS). The number of proteins should have increased because once the sperm cell bursts the intracellular proteins would come out.
I am not familiar with conditions of work with sperm. However I wonder why sperm would "Burst". If I understand well, you increase the osmolarity of the external medium. If this is the case I would not expect many of the sperm proteins to be "released" to the medium as if it were the opposite, i.e. placing sperm in distilled water.
But if it is a matter of disrupting electrostatic interactions, again, increasing the osmotic concentration of the external medium would not necessarily lead to the disruption of electrostatic interactions.
Osmosis is related to sperm releasing water of capturing water
Ionic strength would be related to disruption of electrostatic interactions of "peripheral proteins"
Hello Rafael,
I apologize for my mistake. You are right. However, I am wondering why wouldn't peripheral protein which are bound electrostatically with say GPI linked proteins won't leach out once we disrupt the interactions with elevated salt concentration.
Hi Vipul
I am wondering: perhaps you see what you have to "see" and increasing much the ionic strength may not lead to release more proteins electrostatically bound. I mean perhaps the milder condition is already strong enough.
Alternatively perhaps you are detecting proteins that are enriched in sperm and then the ones released after dirupting electrostatic interactions are hidden (i.e. very diluted in comparison with the abundant ones)
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